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A kit for isothermal detection of bacterial pathogens in respiratory tract infections with enzyme-cleavage probes

A technology of pathogens and respiratory tract, which is applied in the field of reagent kits for the constant temperature detection of bacterial pathogens of respiratory tract infection with enzyme-cutting probes, which can solve the problems of difficulty in achieving sensitivity and specificity

Active Publication Date: 2022-08-05
GENERAL HOSPITAL OF PLA +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, most of the current nucleic acid constant temperature amplification methods are difficult to perform single-tube multiple detection, or the sensitivity and specificity of multiple detection are difficult to achieve the effect of single multiple detection.

Method used

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  • A kit for isothermal detection of bacterial pathogens in respiratory tract infections with enzyme-cleavage probes
  • A kit for isothermal detection of bacterial pathogens in respiratory tract infections with enzyme-cleavage probes
  • A kit for isothermal detection of bacterial pathogens in respiratory tract infections with enzyme-cleavage probes

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1: Determination of the primer probe combination of 12 bacterial disease primary nucleic acid detection of 12 bacterial disease

[0028] 1. Design of single -weight primers and probes

[0029] The present invention has against Golden Polyris (SA), Copper Green Pseudomona (PA), Balman Bacteria (ABA), Pneumonia Crastacilla (KPN), Bachalia (Hi), Purakami Legion (LP), bowel gallery (EC), pneumonia, pneumococcal (SP), malt narrow -eating fake monolm (SME), onion Berk Holde (BC), E. E. E. Cororneya (ECOLI), Star -shaped slaves Bacterial pathogenic gene sequences such as Capillary (NUA) are analyzed. Based on the basic principles of the primer probe design, the design software is used for designing. The combination of the three sets of primers is shown in Table 1.

[0030] Table 1:12 Sequence information of the ranging of bacterial pathogenic programs of the primer

[0031]

[0032]

[0033]

[0034]

[0035]

[0036] 2. Screening of single -weight primers and pr...

Embodiment 2

[0059] Example II: Verification of the testing of the test kit and the sensitivity and specificity of the kit in the present invention

[0060] 1. Preparation kit

[0061] Four nucleic acid reaction liquids, blank control products, and positive control products are packed together, and the product uses instructions are accompanied by the instructions of the enzyme cutting probe in the present invention. Among them, the composition of nucleic acid reaction solution, blank control and positive control products is shown in Table 6 below to detect the fluorescent group marks of the probe of each pathogen.

[0062] Table 6. The ingredients composition of the kit

[0063]

[0064] Table 7. Each probe in the kit The fluorescent base group mark instructions

[0065]

[0066]

[0067] 2. Test 10 cases of clinical sample DNA in the kit in the present invention

[0068] 2.1 The DNA information of the 10 clinical samples used in the test is shown in Table 8 below.

[0069] Table 8. Test...

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Abstract

The present invention provides a kit for the constant temperature detection of bacterial pathogens of respiratory tract infection with enzyme-cleavage probes. The primers and probes of 12 pathogen targets are combined into four triple nucleic acid reaction solutions, and the probes for detecting each bacterial pathogen contain one RNA. Base probes and include 6 isothermal amplification primers. Using the nucleic acid detection kit provided in the present invention, four single-tube multiple constant temperature amplification can be used to detect 12 kinds of bacterial pathogens with high frequency in respiratory tract infections, and the kit is easy to operate, and bedside detection can be realized, and the entire detection time Within 30 minutes, it can quickly and accurately screen a variety of bacterial pathogens in the sample to be tested.

Description

Technical field [0001] The invention involves the field of nucleic acid constant temperature detection technology, and specially involves a kit that specializes in the aquacilids infection of bacterial disease infection with enzyme cutting probe. Background technique [0002] Highness of respiratory infection is common in clinical clinical. Bacterial infections are common causes of respiratory diseases. There are many pathogenic bacteria that cause respiratory bacterial infections. In adult respiratory bacterial infections, golden pylori, copper green single bacteria, and Bowman Berobacteria, pneumonia, albumin, Bacteria, Bacteria, Puomy Legion, Enterprise, Pneumococcus, Pseudomonal Malcaria, Pseudoma, Onion Berk Holde The 12 are the most common. Therefore, it can achieve specific and rapid screening tests for the common bacterial diseases of these 12 respiratory tracts, which facilitates clinicians to diagnose patients in a timely manner and provide appropriate medication treatm...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/689C12Q1/6844C12Q1/04C12Q1/10C12Q1/14C12N15/11C12R1/445C12R1/19C12R1/01C12R1/385C12R1/46C12R1/38C12R1/21C12R1/22
CPCC12Q1/689C12Q1/6844C12Q2600/16C12Q2537/143
Inventor 解立新王凯飞刘利成王华贵蔡金玉唐奇陶毅冯华华王鹏志
Owner GENERAL HOSPITAL OF PLA
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